Quantitative Analysis of Secreted Catecholamines from Chromaffin Cells in Vitro and in Vivo. |
Jun Mo Park, Su Jeong Kim, Kyung Hwa Kwak, Young Hoon Jeon, Woon Yi Baek |
1Department of Anesthesiology and Pain Medicine, Kyungpook National University Hospital, School of Medicine, Korea. webaik@mail.knu.ac.kr 2Department of Medical and and Biological Engineering, Graduate School, Kyungpook National University, Daegu, Korea. |
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Abstract |
BACKGROUND Adrenal medullary transplants into the subarachnoid space have been demonstrated to reduce pain sensitivity. This analgesia most likely results from the release of neuroactive substances, particularly catecholamines and opioid peptides from the transplanted cells into spinal cord. METHODS Isolated bovine chromaffin cells were encapsulated with alginate and poly-L-lysine prior to implantation into rat's subarachnoid space to protect them from host immune system. And then catecholamines from encapsulated chromaffin cells were measured quantitatively in vitro by High Performance Liquid Chromatograph. The animals were randomized into 2 groups, one of which received microencapsulated chromaffin cells and the other empty capsules. The effects of such implants were evaluated on the pain behavior resulting from a chronic constriction injury of the rat sciatic nerve for 30 days. RESULTS Catecholamine concentration in cerebrospinal fluid (CSF) was analyzed. Data (mean SD) are considered significant at P <0.05 (ANOVA for repeated measure and Dunnett's test). Continuous release of catecholamine and met-enkephalin with responsiveness to nicotine stimulation was measured from encapsulated cells in vitro. A significant reduction of allodynic response to acetone evaporation was observed in the animals implanted with cell loaded capsules compared to control animals with empty capsules.
Catecholamine concentration in CSF was higher in the cell loaded capsule group. There were no complications related to implantation. CONCLUSION We found that encapsulated chromaffin cells released continuously catehcolamines and opioids peptides in vitro and in the CSF. Those results may prove chromaffin cell's anagesic effect indirectly. |
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